A cGAL-UAS bipartite expression toolkit for Caenorhabditis elegans sensory neurons.

Animals integrate sensory information from the environment and display various behaviors in response to external stimuli. In Caenorhabditis elegans hermaphrodites, 33 types of sensory neurons are responsible for chemosensation, olfaction, and mechanosensation. However, the functional roles of all sensory neurons have not been systematically studied due to the lack of facile genetic accessibility. A bipartite cGAL-UAS system has been previously developed to study tissue- or cell-specific functions in C. elegans. Here, we report a toolkit of new cGAL drivers that can facilitate the analysis of a vast majority of the 60 sensory neurons in C. elegans hermaphrodites. We generated 37 sensory neuronal cGAL drivers that drive cGAL expression by cell-specific regulatory sequences or intersection of two distinct regulatory regions with overlapping expression (split cGAL). Most cGAL-drivers exhibit expression in single types of cells. We also constructed 28 UAS effectors that allow expression of proteins to perturb or interrogate sensory neurons of choice. This cGAL-UAS sensory neuron toolkit provides a genetic platform to systematically study the functions of C. elegans sensory neurons.

Characteristics of Medical Evacuation by Train in Ukraine, 2022.

Importance: The 2022 war in Ukraine severely affected access to health care for patients in the conflict-affected regions and limited options for medical evacuation. Air transport, a common method of medical evacuation in war zones, was unsafe due to the conflict of 2 modernized military forces that were in possession of aircraft and surface-to-air weapons; therefore, Médecins Sans Frontières, in collaboration with the Ukrainian railway company and Ukrainian health agencies, addressed this by initiating medical evacuation via medically customized trains. Objective: To describe the implementation of medical evacuation trains aimed at improving the access to health care for war-affected patients. Design, Setting, and Participants: This case series describes the remodeling of 2 trains used for medical evacuation in a conflict zone during the war in Ukraine. The study was conducted from March 30 to November 30, 2022. One train had minimal adjustments and could be rapidly deployed to address the most pressing humanitarian needs, while the other underwent major structural modifications to provide intensive care capacity. The report details the medical capabilities of the trains, the organization of referrals, and operational challenges encountered. Additionally, it includes a case series on the characteristics of patients transported in the initial 8 months, based on routinely collected programmatic descriptive data of all patients transported by the medical trains. Results: In 8 months, 2481 patients (male-female ratio, 1.07; male, 1136 [46%]; female 1058 [43%]; missing data, 287 [12%]; median age, 63 years [range, 0-98 years]) were evacuated from 11 cities near the Ukrainian conflict frontline to safer areas. Initially, the trains predominantly evacuated trauma patients, but over the course of the war, the patient characteristics changed with more medical and nonacute conditions, and fewer trauma patients. The main reason for entry into the intensive care unit train carriage was for close monitoring and observation, and the main interventions performed were primarily for respiratory failure. Conclusions and Relevance: The findings of this study suggest that medical evacuation in a war zone by converted trains is possible and can improve access to health care for war-affected patients. The presence of intensive care capacity on board allows for transport of more severely ill or injured individuals. However, the target population should not be limited to trauma patients, as health care institutions affected host a much broader population whose needs and urgency for evacuation may change over time.

HIV silencing and cell survival signatures in infected T cell reservoirs.

Rare CD4 T cells that contain HIV under antiretroviral therapy represent an important barrier to HIV cure1-3, but the infeasibility of isolating and characterizing these cells in their natural state has led to uncertainty about whether they possess distinctive attributes that HIV cure-directed therapies might exploit. Here we address this challenge using a microfluidic technology that isolates the transcriptomes of HIV-infected cells based solely on the detection of HIV DNA. HIV-DNA+ memory CD4 T cells in the blood from people receiving antiretroviral therapy showed inhibition of six transcriptomic pathways, including death receptor signalling, necroptosis signalling and antiproliferative Gα12/13 signalling. Moreover, two groups of genes identified by network co-expression analysis were significantly associated with HIV-DNA+ cells. These genes (n = 145) accounted for just 0.81% of the measured transcriptome and included negative regulators of HIV transcription that were higher in HIV-DNA+ cells, positive regulators of HIV transcription that were lower in HIV-DNA+ cells, and other genes involved in RNA processing, negative regulation of mRNA translation, and regulation of cell state and fate. These findings reveal that HIV-infected memory CD4 T cells under antiretroviral therapy are a distinctive population with host gene expression patterns that favour HIV silencing, cell survival and cell proliferation, with important implications for the development of HIV cure strategies.

Neck Swelling in a Critically Ill Patient With COVID-19-Related Acute Respiratory Distress Syndrome on Venovenous Extracorporeal Membrane Oxygenation: Consider the Differential.

Neck swelling during venovenous extracorporeal membrane oxygenation (VV-ECMO) usually heralds the development of potentially serious complications, including superior vena cava (SVC) syndrome, hematoma, and/or angioedema. In this case report, we describe a 43-year-old male patient who had received VV-ECMO support for the coronavirus disease 2019 (COVID-19) acute respiratory distress syndrome. During his hospitalization, he developed acute onset of neck swelling after two weeks of VV-ECMO and two days after a tracheostomy. Clinical examination and investigations were performed to exclude ECMO-related SVC syndrome and tracheostomy-related complications. Consequently, it was discovered the patient had developed COVID-19-related subacute thyroiditis with enlargement of both thyroid glands. Conservative management, including the use of continued glucocorticoids, raising the head of the bed, and observing for complications of thyroiditis, was undertaken. Eventually, this patient's neck swelling resolved on its own, and he was eventually decannulated from ECMO several weeks later. Our case report highlights the differential diagnosis of neck swelling during VV-ECMO and considers the evaluation of different etiologies.

Insulin/IGF signaling regulates presynaptic glutamate release in aversive olfactory learning.

Insulin/insulin-like growth factor (IGF) receptor signaling (IIS) supports context-dependent learning in vertebrates and invertebrates. Here, we identify cell-specific mechanisms of IIS that integrate sensory information with food context to drive synaptic plasticity and learning. In the nematode Caenorhabditis elegans, pairing food deprivation with an odor such as butanone suppresses attraction to that odor. We find that aversive olfactory learning requires the insulin receptor substrate (IRS) protein IST-1 and atypical signaling through the insulin/IGF-1 receptor DAF-2. Cell-specific knockout and rescue demonstrate that DAF-2 acts in the AWCON sensory neuron, which detects butanone, and that learning preferentially depends upon the axonally localized DAF-2c isoform. Acute food deprivation increases DAF-2 levels in AWCON post-transcriptionally through an insulin- and insulin receptor substrate-1 (ist-1)-dependent process. Aversive learning alters the synaptic output of AWCON by suppressing odor-regulated glutamate release in wild-type animals, but not in ist-1 mutants, suggesting that axonal insulin signaling regulates synaptic transmission to support aversive memory.

Esophageal Balloon-Directed Ventilator Management for Postpneumonectomy Acute Respiratory Distress Syndrome.

OBJECTIVE: Postpneumonectomy patients may develop acute respiratory distress syndrome (ARDS). There is a paucity of data regarding the optimal management of mechanical ventilation for postpneumonectomy patients. Esophageal balloon pressure monitoring has been used in traditional ARDS patients to set positive end-expiratory pressure (PEEP) and minimize transpulmonary driving pressure (ΔP L), but its clinical use has not been previously described nor validated in postpneumonectomy patients. The primary objective of this report was to describe the potential clinical application of esophageal pressure monitoring to manage the postpneumonectomy patient with ARDS. DESIGN: Case report. Setting. Surgical intensive care unit (ICU) of a university-affiliated teaching hospital. Patient. A 28-year-old patient was involved in a motor vehicle collision, with a right main bronchus injury, that required a right-sided pneumonectomy to stabilize his condition. In the perioperative phase, they subsequently developed ventilator-associated pneumonia, significant cumulative positive fluid balance, and ARDS. Interventions. Prone positioning and neuromuscular blockade were initiated. An esophageal balloon was inserted to direct ventilator management. Measurements and Main Results. V T was kept around 3.6 mL/kg PBW, ΔP L at ≤14 cm H2O, and plateau pressure at ≤30 cm H2O. Lung compliance was measured to be 37 mL/cm H2O. PEEP was optimized to maintain end-inspiratory transpulmonary pressure (P L) < 15 cm H2O, and end-expiratory P L between 0 and 5 cm H2O. The maximal ΔP L was measured to be 11 cm H2O during the care of this patient. The patient improved with esophageal balloon-directed ventilator management and was eventually liberated from mechanical ventilation. CONCLUSIONS: The optimal targets for V T remain unknown in the postpneumonectomy patient. However, postpneumonectomy patients with ARDS may potentially benefit from very low V T and optimization of PEEP. We demonstrate the application of esophageal balloon pressure monitoring that clinicians could potentially use to limit injurious ventilation and improve outcomes in postpneumonectomy patients with ARDS. However, esophageal balloon pressure monitoring has not been extensively validated in this patient population.

Anti-Ebola therapy for patients with Ebola virus disease: a systematic review.

BACKGROUND: Management of Ebola virus disease (EVD) has historically focused on infection prevention, case detection and supportive care. Several specific anti-Ebola therapies have been investigated, including during the 2014-2016 West African outbreak. Our objective was to conduct a systematic review of the effect of anti-Ebola virus therapies on clinical outcomes to guide their potential use and future evaluation. METHODS: We searched PubMed, EMBASE, Global Health, Cochrane Library, African Index Medicus, WHOLIS (inception-9 April 2018), and trial registries for observational studies or clinical trials, in any language, that enrolled patients with confirmed EVD who received therapy targeting Ebola virus and reported on mortality, symptom duration, or adverse effects. RESULTS: From 11,257 citations and registered trials, we reviewed 55 full-text citations, of which 35 met eligibility criteria (1 randomized clinical trial (RCT), 8 non-randomized comparative studies, 9 case series and 17 case reports) and collectively examined 21 anti-Ebola virus agents. The 31 studies performed during the West African outbreak reported on 4.8% (1377/28616) of all patients with Ebola. The only RCT enrolled 72 patients (0.25% of all patients with Ebola) and compared the monoclonal antibody ZMapp vs. standard care (mortality, 22% vs. 37%; 95% confidence interval for risk difference, - 36 to 7%). Studies of convalescent plasma, interferon-ß-1a, favipiravir, brincidofovir, artesunate-amodiaquine and TKM-130803 were associated with at least moderate risk of bias. CONCLUSIONS: Research evaluating anti-Ebola virus agents has reached very few patients with EVD, and inferences are limited by non-randomized study designs. ZMapp has the most promising treatment signal.

Caring for Critically Ill Patients in Humanitarian Settings.

Critical care medicine is far from the first medical field to come to mind when humanitarian action is mentioned, yet both critical care and humanitarian action share a fundamental purpose to save the lives and ease the suffering of people caught in acute crises. Critically ill children and adults will be present regardless of resource limitations and irrespective of geography, regional or cultural contexts, insecurity, or socioeconomic status, and they may be even more prevalent in a humanitarian crisis. Critical care is not limited to the walls of a hospital, and all hospitals will have critically ill patients regardless of designating a specific ward an ICU. Regular and consistent consideration of critical care principles in humanitarian settings provides crucial guidance to intensivists and nonintensivists alike. A multidisciplinary, systematic approach to patient care that encourages critical thinking, checklists that encourage communication among team members, and context-specific critical care rapid response teams are examples of critical care constructs that can provide high-quality critical care in all environments. Promoting critical care principles conveys the message that critical care is an integral part of health care and should be accessible to all, no matter the setting. These principles can be effectively adopted in humanitarian settings by normalizing them to everyday clinical practice. Equally, core humanitarian principles-dignity, accountability, impartiality, neutrality-can be applied to critical care. Applying principles of critical care in a context-specific manner and applying humanitarian principles to critical care can improve the quality of patient care and transcend barriers to resource limitations.

The Small Molecule NLRP3 Inflammasome Inhibitor MCC950 Does Not Alter Wound Healing in Obese Mice.

The incidence of chronic wounds is escalating, and the associated healing process is especially problematic in an aging population with increased morbidity. Targeting increased inflammation in chronic wounds is a promising but challenging therapeutic strategy. Indeed, inflammation and especially macrophages are required for wound healing. As the NLRP3 inflammasome has been implicated with various other inflammatory diseases, in this study, we used MCC950-a selective NLRP3 small molecule inhibitor-on murine models of both acute and chronic wounds. This molecule, while tested for other inflammatory conditions, has never been investigated to reduce topical inflammation driving chronic wounds. We found that there were no significant differences when the treatment was applied either topically or orally in wild-type C57Bl/6 mice and that it even impaired wound healing in obese mice. The treatment was also unable to improve re-epithelialisation or angiogenesis, which are both required for the closure of wounds. We are inclined to believe that MCC950 may inhibit the closure of chronic wounds and that it does not alter wound-associated macrophage polarisation.

Accelerated Endothelial to Mesenchymal Transition Increased Fibrosis via Deleting Notch Signaling in Wound Vasculature.

Skin wound healing in adults is characterized by a peak of angiogenesis followed by regression of the excessive vasculature in parallel with collagen deposition and fibrosis in the wound. We hypothesized that regressing vessels in healing wounds were in fact entering an endothelial to mesenchymal transition contributing to scarring. Using vascular-specific fate tracking (Cdh5-creERt2/ROSA-YFP mice), full-thickness excisional wounds were analyzed to reveal a time-dependent transition from endothelial phenotype characterized by vascular endothelial-cadherin, CD31, and CD34 toward a mesenchymal phenotype characterized by alpha-smooth muscle actin and fibroblast-specific protein 1 expression. We next conditionally ablated RBPJ in the vasculature (Rbpjfl/fl/Cdh5-creERt2ROSA-YFP) to evaluate the role of canonical Notch signaling in this process. Endothelial to mesenchymal transition was clearly accelerated after the loss of Notch signaling within the vasculature. The acceleration of endothelial to mesenchymal transition resulted in delayed wound healing, increased fibrosis, and extensive scar tissue formation, with the rapid loss of key endothelial genes and proteins and upregulation of mesenchymal protein expression (alpha-smooth muscle actin and fibroblast-specific protein 1) in vessels. Our findings here uncover a cellular contributor to skin wound scarring through the process of endothelial to mesenchymal transition in skin wounds and demonstrate the importance of Notch signaling in regulating this critical process during healing.

Mesenchymal stem/stromal cells enhance engraftment, vasculogenic and pro-angiogenic activities of endothelial colony forming cells in immunocompetent hosts.

The clinical use of endothelial colony forming cells (ECFC) is hampered by their restricted engraftment. We aimed to assess engraftment, vasculogenic and pro-angiogenic activities of ECFC in immunocompetent (C57BL/6: WT) or immunodeficient (rag1 -/- C57BL/6: Rag1) mice. In addition, the impact of host immune system was investigated where ECFC were co-implanted with mesenchymal stem/stromal cells (MSC) from adult bone marrow (AdBM-MSC), fetal bone marrow (fBM-MSC), fetal placental (fPL-MSC), or maternal placental (MPL-MSC). Transplantation of ECFCs in Matrigel plugs resulted in less cell engraftment in WT mice compared to Rag1 mice. Co-implantation with different MSCs resulted in a significant increase in cell engraftment up to 9 fold in WT mice reaching levels of engraftment observed when using ECFCs alone in Rag1 mice but well below levels of engraftment with MSC-ECFC combination in Rag1 recipients. Furthermore, MSCs did not reduce murine splenic T cell proliferation in response to ECFCs in vitro. ECFCs enhanced the murine neo-vascularization through paracrine effect, but with no difference between Rag1 and WT mice. In conclusions, the host adaptive immune system affects the engraftment of ECFCs. MSC co-implantation improves ECFC engraftment and function even in immunocompetent hosts mostly through non-immune mechanisms.

Multiple Origins of Virus Persistence during Natural Control of HIV Infection.

Targeted HIV cure strategies require definition of the mechanisms that maintain the virus. Here, we tracked HIV replication and the persistence of infected CD4 T cells in individuals with natural virologic control by sequencing viruses, T cell receptor genes, HIV integration sites, and cellular transcriptomes. Our results revealed three mechanisms of HIV persistence operating within distinct anatomic and functional compartments. In lymph node, we detected viruses with genetic and transcriptional attributes of active replication in both T follicular helper (TFH) cells and non-TFH memory cells. In blood, we detected inducible proviruses of archival origin among highly differentiated, clonally expanded cells. Linking the lymph node and blood was a small population of circulating cells harboring inducible proviruses of recent origin. Thus, HIV replication in lymphoid tissue, clonal expansion of infected cells, and recirculation of recently infected cells act together to maintain the virus in HIV controllers despite effective antiviral immunity.

Viraemia and Ebola virus secretion in survivors of Ebola virus disease in Sierra Leone: a cross-sectional cohort study.

BACKGROUND: In survivors of Ebola virus disease, clinical sequelae including uveitis, arthralgia, and fatigue are common and necessitate systematic follow-up. However, the infection risk to health-care providers is poorly defined. Here we report Ebola virus RT-PCR data for body site and fluid samples from a large cohort of Ebola virus survivors at clinic follow-up. METHODS: In this cross-sectional cohort study, consecutive survivors of Ebola virus disease attending Kerry Town survivor clinic (Freetown, Sierra Leone), who had been discharged from the Kerry Town Ebola treatment unit, were invited to participate. We collected and tested axillary, blood, conjunctival, forehead, mouth, rectal, semen, urine, and vaginal specimens for presence of Ebola virus using RT-PCR. We regarded samples to be positive for Ebola virus disease if the cycle threshold was 40 or lower. We collected demographic data from survivors of their age, sex, time since discharge from the treatment unit, and length of acute admission in the Ebola treatment unit using anonymised standard forms. FINDINGS: Between April 2, and June 16, 2015, of 151 survivors of Ebola virus disease invited to participate, 112 (74%) provided consent. The median age of participants was 21·5 years (IQR 14-31·5) with 34 (30%) participants younger than 16 years. 50 (45%) of 112 participants were male. We tested a total of 555 specimens: 103 from the axilla, 93 from blood, 92 from conjunctiva, 54 from forehead, 105 from mouth, 17 from the rectum, one from semen, 69 from urine, and 21 from the vagina. The median time from Ebola treatment unit discharge to specimen collection was 142 days (IQR 127-159). 15 participants had a total of 74 swabs taken less than 100 days from discharge. The semen sample from one participant tested positive for Ebola virus at 114 days after discharge from the treatment unit; specimens taken from the axilla, blood, conjunctiva, forehead, mouth, rectum, and urine of the same participant tested negative. All specimens from the other 111 participants tested negative. INTERPRETATION: Patients recovering from Ebola virus disease who do not meet the case definition for acute disease pose a low infection risk to health-care providers 6 weeks after clearance of viraemia. Personal protective equipment after this time might be limited to standard barrier precautions, unless contact with fluids from sanctuary sites is envisaged. FUNDING: Save the Children International, Public Health England.

Empiric intravenous fluid and electrolyte therapy in patients with Ebola virus disease.

The role of intravenous fluid and electrolyte replacement is increasingly recognised in the clinical management of Ebola virus disease. However, outbreaks typically occur in resource-limited settings where electrolyte measurement may be initially unavailable. Here we highlight potential strategies for empiric fluid and electrolyte therapy based on experience from Sierra Leone.

High Performance Flexible Actuator of Urchin-Like ZnO Nanostructure/Polyvinylenefluoride Hybrid Thin Film with Graphene Electrodes for Acoustic Generator and Analyzer.

A bass frequency response enhanced flexible polyvinylidene fluoride (PVDF) based thin film acoustic actuator is successfully fabricated. High concentrations of various zinc oxide (ZnO) is embedded in PVDF matrix, enhancing the ß phase content and the dielectric property of the composite thin film. ZnO acts as a nucleation agent for the crystallization of PVDF. A chemical vapor deposition grown graphene is used as electrodes, enabling high electron mobility for the distortion free acoustic signals. The frequency response of the fabricated acoustic actuator is studied as a function of the film thickness and filler content. The optimized film has a thickness of 80 µm with 30 wt% filler content and shows 72% and 42% frequency response enhancement in bass and midrange compared to the commercial PVDF, respectively. Also, the total harmonic distortion decreases to 82% and 74% in the bass and midrange regions, respectively. Furthermore, the composite film shows a promising potential for microphone applications. Most of all, it is demonstrated that acoustic actuator performance is strongly influenced by degree of PVDF crystalline.

Highly Omnidirectional and Frequency Controllable Carbon/Polyaniline-based 2D and 3D Monopole Antenna.

Highly omnidirectional and frequency controllable carbon/polyaniline (C/PANI)-based, two- (2D) and three-dimensional (3D) monopole antennas were fabricated using screen-printing and a one-step, dimensionally confined hydrothermal strategy, respectively. Solvated C/PANI was synthesized by low-temperature interfacial polymerization, during which strong π-π interactions between graphene and the quinoid rings of PANI resulted in an expanded PANI conformation with enhanced crystallinity and improved mechanical and electrical properties. Compared to antennas composed of pristine carbon or PANI-based 2D monopole structures, 2D monopole antennas composed of this enhanced hybrid material were highly efficient and amenable to high-frequency, omnidirectional electromagnetic waves. The mean frequency of C/PANI fiber-based 3D monopole antennas could be controlled by simply cutting and stretching the antenna. These antennas attained high peak gain (3.60 dBi), high directivity (3.91 dBi) and radiation efficiency (92.12%) relative to 2D monopole antenna. These improvements were attributed the high packing density and aspect ratios of C/PANI fibers and the removal of the flexible substrate. This approach offers a valuable and promising tool for producing highly omnidirectional and frequency-controllable, carbon-based monopole antennas for use in wireless networking communications on industrial, scientific, and medical (ISM) bands.

Impact of a Two-step Emergency Department Triage Model with START, then CTAS, on Patient Flow During a Simulated Mass-casualty Incident.

INTRODUCTION: A high influx of patients during a mass-casualty incident (MCI) may disrupt patient flow in an already overcrowded emergency department (ED) that is functioning beyond its operating capacity. This pilot study examined the impact of a two-step ED triage model using Simple Triage and Rapid Treatment (START) for pre-triage, followed by triage with the Canadian Triage and Acuity Scale (CTAS), on patient flow during a MCI simulation exercise. Hypothesis/Problem It was hypothesized that there would be no difference in time intervals nor patient volumes at each patient-flow milestone. METHODS: Physicians and nurses participated in a computer-based tabletop disaster simulation exercise. Physicians were randomized into the intervention group using START, then CTAS, or the control group using START alone. Patient-flow milestones including time intervals and patient volumes from ED arrival to triage, ED arrival to bed assignment, ED arrival to physician assessment, and ED arrival to disposition decision were compared. Triage accuracy was compared for secondary purposes. RESULTS: There were no significant differences in the time interval from ED arrival to triage (mean difference 108 seconds; 95% CI, -353 to 596 seconds; P=1.0), ED arrival to bed assignment (mean difference 362 seconds; 95% CI, -1,269 to 545 seconds; P=1.0), ED arrival to physician assessment (mean difference 31 seconds; 95% CI, -1,104 to 348 seconds; P=0.92), and ED arrival to disposition decision (mean difference 175 seconds; 95% CI, -1,650 to 1,300 seconds; P=1.0) between the two groups. There were no significant differences in the volume of patients to be triaged (32% vs 34%; 95% CI for the difference -16% to 21%; P=1.0), assigned a bed (16% vs 21%; 95% CI for the difference -11% to 20%; P=1.0), assessed by a physician (20% vs 22%; 95% CI for the difference -14% to 19%; P=1.0), and with a disposition decision (20% vs 9%; 95% CI for the difference -25% to 4%; P=.34) between the two groups. The accuracy of triage was similar in both groups (57% vs 70%; 95% CI for the difference -15% to 41%; P=.46). CONCLUSION: Experienced triage nurses were able to apply CTAS effectively during a MCI simulation exercise. A two-step ED triage model using START, then CTAS, had similar patient flow and triage accuracy when compared to START alone.